恩诺沙星诱导溶藻弧菌耐药和交叉耐药的研究  

Analysis of Drug-Resistance and Cross Resistance in Vitro of Vibrio alginolyticus Induced by Enrofloxacin

作　　者：张凯[1,2] 张明明 谢曦 王蓉 胡文婷[1] ZHANG Kai;ZHANG Mingming;XIE Xi;WANG Rong;HU Wenting(Key Laboratory of Tropical Biological Resources, Ministry of Education, College of Life Sciences and Pharmacy, Hainan University, Haikou 570228, China;Hainan Kangzhi Pharmaceutical Co., Ltd., Haikou 570311, China)

机构地区：[1]海南大学药学院,热带生物资源教育部重点实验室,海南海口570228 [2]康芝药业股份有限公司,海南海口570311

出　　处：《水产科学》2022年第3期452-458,共7页Fisheries Science

基　　金：海南省自然科学基金青年基金资助项目(319QN160);海南大学科研启动基金项目(KYQD(ZR)19108).

摘　　要：自收集于海南海水养殖区域内的溶藻弧菌中筛选恩诺沙星敏感株,在含亚抑菌质量浓度恩诺沙星的培养基上进行质量浓度递增法体外诱导,探究耐药性的发生状况,比较诱导耐药前后诱导菌对诱导药物和6种非诱导药物的最小抑菌质量浓度,并在不含药物的培养基上连续传代考察诱导后的耐药株遗传稳定性。试验结果表明,30株养殖源溶藻弧菌中筛选出24株恩诺沙星敏感株;经30代体外诱导,敏感株均稳定获得了恩诺沙星耐药,最小抑菌质量浓度为诱导前的64~1024倍;诱导菌株对非诱导的喹诺酮类药物诺氟沙星和环丙沙星产生明显的交叉耐药,大多数菌株对四环素表现出了交叉耐药,对氟苯尼考和磺胺嘧啶表现出一定的耐药性增强,对硫酸新霉素的耐药性未发生明显变化。14株溶藻弧菌诱导耐药株的交叉耐药率达到50%以上,其中菌株HNVA21的交叉耐药比最高,达83.3%。耐药子代在无药物培养基中连续传代20次后,21株最小抑菌质量浓度保持不变,耐药性稳定,3株耐药菌株耐药性下降为诱导后的1/2。亚抑菌质量浓度恩诺沙星可诱导溶藻弧菌产生耐药,并对多种非诱导药物产生交叉耐药,获得的耐药株遗传稳定性较好。Enrofloxacin-sensitive Vibrio alginolyticus strains collected from the marine aquaculture area in Hainan province were induced by concentration-increasing method on the medium containing sub-inhibitory concentration of enrofloxacin in vitro to evaluate the occurrence of drug resistance.The minimal inhibitory concentration(MIC)of the induced bacterium to induced drugs and 6 non induced drugs was compared before and after drug resistance induction,and the genetic stability of the induced drug resistance strains was investigated on the media without drugs.The results showed that 24 enrofloxacin-sensitive strains were screened out of 30 strains of V.alginolyticus after 30 generations of induction in vitro.The sensitive strains were all resistant to enrofloxacin,and the MIC was 64—1024 times as large as before induction.The induced strains had severe cross-resistance to the non-induced quinolone drugs ciprofloxacin and norfloxacin,increase in resistance to tetracycline,a certain degree of enhancement to florfenicol and sulfadiazine,and no significant changes in sulfadiazine resistance.The cross resistance rates of 14 strains were over 50%,and that of HNVA21 was 83.3%.After 20 successive passages of drug-resistant progeny,the MIC value of 21 strains remained unchanged,and 3 strains were decreased by 1/2,with relatively stable drug resistance.Enrofloxacin led to induce drug resistance of V.alginolyticus with secondary inhibitory concentration,and cross-resistance to a variety of non-induced drugs,with good genetic stability of the resistant strains.

关 键 词：溶藻弧菌 耐药 恩诺沙星 体外诱导 

分 类 号：S941.42[农业科学—水产养殖]